Productive Infection of Bovine Papillomavirus Type 2 in the Urothelial Cells of Naturally Occurring Urinary Bladder Tumors in Cattle and Water Buffaloes
Background Papillomaviruses (PVs) are highly epitheliotropic as they usually establish productive infections within squamous epithelia of the skin, the anogenital tract and the oral cavity. In this study, early (E) and late (L) protein expression of bovine papillomavirus type 2 (BPV-2) in the urothelium of the urinary bladder is described in cows and water buffaloes suffering from naturally occurring papillomavirus-associated urothelial bladder tumors. Methods and Findings E5 protein, the major oncoprotein of the BPV-2, was detected in all tumors. L1 DNA was amplified by PCR, cloned and sequenced and confirmed to be L1 DNA. The major capsid protein, L1, believed to be only expressed in productive papillomavirus infection was detected by Western blot analysis. Immunohistochemical investigations confirmed the presence of L1 protein both in the cytoplasm and nuclei of cells of the neoplastic urothelium. Finally, the early protein E2, required for viral DNA replication and known to be a pivotal factor for both productive and persistent infection, was detected by Western blot and immunohistochemically. Electron microscopic investigations detected electron dense particles, the shape and size of which are consistent with submicroscopic features of viral particles, in nuclei of neoplastic urothelium. Conclusion This study shows that both active and productive infections by BPV-2 in the urothelium of the bovine and bubaline urinary bladder can occur in vivo.
References
[1]
Moody CA, Laimins LA (2010) Human papillomavirus oncoproteins: pathways to transformation. Nat Rev Cancer 10: 550–556.
[2]
IARC (2007) IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Vol 90, Human Papillomaviruses. Lyon, France: IARC Press.
[3]
Zhu W, Dong J, Shimizu E, Hatama S, Kadota K, et al. (2012) Characterization of novel bovine papillomavirus type 12 (BPV-12) causing epithelial papilloma. Arch Virol 157: 17–27.
[4]
Lunardi M, Alfieri AA, Otonel RA, de Alcantara BK, Rodrigues WB, et al. (2013) Genetic characterization of a novel bovine papillomavirus member of the Deltapapillomavirus genus. Vet Microbiol 162: 207–213.
[5]
Shafti-Keramat S, Schellenbacher C, Handisurya A, Christensen N, Reininger B, et al. (2009) Bovine papillomavirus type 1 (BPV1) and BPV2 are closely related serotypes. Virology 393: 1–6.
[6]
de Villiers ET, Fauquet C, Broker TR, Bernard HU, zur Hausen H (2004) Classification of papillomaviruses. Virology 324: 17–27.
[7]
Graham SV (2006) Late events in the life cycle of human papillomavirus. In: Campo MS, editor. Papillomavirus Research - from Natural History to Vaccines and Beyond, pp 193–211, Norfolk, UK: Caister Academic Press.
[8]
Roperto S, Comazzi S, Ciusani E, Paolini F, Borzacchiello G, et al. (2011) PBMCs are additional sites of productive infection of bovine papillomavirus type 2. J Gen Virol 92: 1787–1794.
[9]
Roperto S, Borzacchiello G, Esposito I, Riccardi M, Urraro C, et al. (2012) Productive infection of bovine papillomavirus type 2 in the placenta of pregnant cows affected with urinary bladder tumors. PLoS One 7 (3) e33569 doi:10.1371/journal.pone.0033569.
[10]
Borzacchiello G, Russo V, Gentile F, Roperto F, Venuti A, et al. (2006) Bovine papillomavirus E5 oncoprotein binds to the activated form of the platelet-derived growth factor beta receptor in naturally occurring bovine urinary bladder tumours. Oncogene 25: 1251–1260.
[11]
Roperto S, De Tullio R, Raso C, Stifanese R, Russo V, et al. (2010) Calpain3 is expressed in a proteolitically active form in papillomavirus-associated urothelial tumors of the urinary bladder in cattle. PLoS One 5 (4) e10299 doi: 10.1371/journal.pone.0010299.
[12]
Muller M, Jacob Y, Jones L, Weiss A, Brino L, et al. (2012) Large scale genotype comparison of human papillomavirus E2-host interaction networks provides new insights for E2 molecular functions. PLoS Pathog 8 (6) e1002761 doi:10.1371/journal.ppat.1002761.
[13]
Johansson C, Somberg M, Li X, Backstr？m Winquist E, Fay J, et al. (2012) HPV-16 E2 contributes to induction of HPV-16 late gene expression by inhibiting early polyadenylation. EMBO J 31: 3212–3227.
[14]
Roperto S, Brun R, Paolini F, Urraro C, Russo V, et al. (2008) Detection of bovine papillomavirus type 2 in the peripheral blood of cattle with urinary bladder tumours: a possible biological role. J Gen Virol 89: 3027–3033.
[15]
Campo MS (2006) Bovine papillomavirus: old system, new lessons? In: Campo MS, editor. Papillomavirus Research - from Natural History to Vaccines and Beyond, pp 373–387, Norfolk, UK:Caister Academic Press.
[16]
Pathania S, Dhama K, Saikumar G, Shahi S, Somvanshi R (2012) Detection and quantification of bovine papilloma virus type 2 (BPV-2) by Real-PCR in urine and urinary bladder lesions in enzootic bovine haematuria (EBH)-affected cows. Transbound Emerg Dis 59: 79–84.
[17]
Somvanshi R (2011) Papillomatosis in buffaloes: a less-known disease. Transbound Emerg Dis 58: 327.332.
[18]
Somvanshi R, Pathania S, Nagarajan N, Pangty K, Kamar P (2012) Pathological study of non-neoplastic urinary bladder lesions in cattle and buffaloes: a preliminary report. Trop Anim Health Prod 44: 855–861.
[19]
Roperto S, Russo V, Ozkul A, Sepici-Dincel A, Maiolino P, et al. (2013) Bovine papillomavirus type 2 infects the urinary bladder of water buffalo (Bubalus bubalis) and plays a crucial role in the bubaline urothelial carcinogenesis. J Gen Virol 94: 403–408.
[20]
Shigehara K, Sasagawa T, Kawaguchi S, Nakashima T, Shimamura M, et al. (2011) Etiologic role of human papillomavirus infection in bladder carcinoma. Cancer 117: 2067–2076.
[21]
Stocco dos Santos RC, Lindsey CJ, Ferraz OP, Pinto JR, Mirandola RS, et al. (1998) Bovine papillomavirus transmission and chromosomal aberrations: an experimental model. J Gen Virol 79: 2127–35.
[22]
Roperto S, Borzacchiello G, Brun R, Leonardi L, Maiolino P, et al. (2010) Review of bovine urothelial tumours and tumour-like lesions of the urinary bladder. J Comp Pathol 142: 95–108.
[23]
Griesser H, Sander H, Walczak C, Hilfrich RA (2009) HPV vaccine protein L1 predicts disease outcome of high-risk HPV+ early squamous dysplastic lesions. Am J Clin Pathol 132: 840–845.
[24]
Bodily J, Laimins LA (2011) Persistence of human papillomavirus infection: keys to malignant progression. Trends in Microbiol 19: 33–39.
[25]
Middleton K, Peh W, Southern S, Griffin H, Sotlar K, et al. (2003) Organization of human papillomavirus productive cycle during neoplastic progression provides a basis for selection of diagnostic markers. J Virol 77: 10186–10201.
[26]
zur Hausen H (2002) Papillomavirus and cancer: from basic studies to clinical application. Nat Rev Cancer 2: 342–350.
[27]
Pett M, Coleman N (2007) Integration of high-risk human papillomavirus: a key event in cervical carcinogenesis? J Pathol 212: 356–367.
[28]
Doorbar J (2007) Papillomavirus life cycle organization and biomarker selection. Dis Markers 23: 297–313.
[29]
H？fner N, Driesch C, Gajda M, Jansen L, Kirchmayr R, et al. (2008) Integration of the HPV16 genome does not invariably result in high levels of viral oncogene transcripts. 27: 1610–1617.
[30]
Das Ghosh D, Bhattacharrjee B, Sen S, Premi L, Mukhopadhyay I, et al. (2012) Some novel insights on HPV16 related cervical cancer pathogenesis based on analyses of LCR methylation, viral load, E7 and E2/E4 expressions. PLoS One 7 (9) e44678 doi:10.1371/journal.pone.0044678.
[31]
Xiao W, Bian M, Ma L, Liu J, Chen Y, et al. (2010) Immunochemical analysis of human papillomavirus L1 capsid protein in liquid-based cytology samples from cervical lesions. Acta Cytol 54: 661–667.
[32]
Yemelyanova A, Gravitt PE, Ronnett BM, Rositch AF, Ogurtsova A, et al. (2013) Immunohistochemical detection of human papillomavirus capsid proteins L1 and L2 in squamous intraepithelial lesions: potential utility in diagnosis and management. Mod Pathol 26: 268–274.
[33]
Sarmadi S, Izadi-mood N, Pourlashkari M, Yarandi F, Sanii S (2010) HPV L1 capsid protein expression in squamous intraepithelial lesions of cervix uteri and its relevance to disease outcome. Arch Gynecol Obstet 285: 779–784.
[34]
Hernandez J, Elahi A, Siegel E, Coppola D, Riggs B, et al. (2011) HPV L1 capsid protein detection and progression of anal squamous neoplasia. Am J Clin Pathol 135: 436–441.
[35]
Melsheimer P, Kaul S, Dobeck S, Bastert G (2003) Immunocytochemical detection of HPV high-risk type L1 capsid proteins in LSIL and HSIL as compared with detection of HPV L1 DNA. Acta Cytol 47: 124–128.
[36]
Shigehara K, Sasagawa T, Doorbar J, Kawaguchi S, Kobori Y, et al. (2011) Etiological role of human papillomavirus infection for inverted papilloma of the bladder. J Med Virol 83: 277–285.
[37]
Campo MS, Jarrett WFH, Barron RJ, O'Neil BW, Smith KT (1992) Association of bovine papillomavirus type 2 and bracken fern with bladder cancer in cattle. Cancer Res 53: 1–7.
[38]
Wosiaki SR, Claus MP, Alfieri AL, Alfieri AA (2005) Bovine papillomavirus type 2 detection in the urinary bladder of cattle with chronic enzootic haematuria. Mem Inst Oswaldo Cruz 101: 635–638.
[39]
Morgan IM, Donaldson MM (2006) The papillomavirus transcription/replication factor E2: structure, function, cancer and therapy. In: Campo MS, editor. Papillomavirus Research - from Natural History to Vaccines and Beyond, pp 73–82, Norfolk, UK:Caister Academic Press.
[40]
Campo MS (2002) Animal model of papillomavirus pathogenesis. Virus Res 89: 249–261.
[41]
Alvarez J, Pokomandy ADE, Rouleau D, Ghattas G, Vezina S, et al. (2010) Episomal and integrated human papillomavirus type 16 loads and anal intraepithelial neoplasia in HIV-seropositive men. AIDS 24: 2355–2363.
[42]
Modis Y, Trus BI, Harrison SC (2001) Atomic model of the papillomavirus capsid. EMBO J 21: 4754–4762.
[43]
Roperto S, Di Guardo G, Leonardi L, Pagnini U, Manco E, et al. (2012) Bacterial isolates from the urine of cattle affected by urothelial tumors of the urinary bladder. Res Vet Sci 93: 1361–1366.
[44]
Peek RM, Mohla S, DuBois RN (2005) Inflammation in the genesis and perpetuation of cancer: summary and recommendations from a National Cancer Institute-sponsored meeting. Cancer Res 65: 8583–8587.
